Abcam Ad

Author Topic: 350um THICK sections, Lectin histochemistry NOT immunohisto!  (Read 2977 times)

0 Members and 1 Guest are viewing this topic.

Offline tara

  • Newbie
  • *
  • Posts: 2
350um THICK sections, Lectin histochemistry NOT immunohisto!
« on: October 07, 2009, 12:09:02 PM »
Hi,
I cant seem to find much information on lectin Histochemical staining.  Im using Vector FL-1351.  And i could do with any suggestions regarding concentrations, incubation times etc
So far all i have come up with is i need an inhibitory sugar as a control, a possible 1hr incubation at RT and a possible 2 -20ug concentration range.

I've never done histochemistry before, the concept is unusual, i would prefer to have a secondary. Do i need to do washes??  Do i need Triton X as its a thicker slice.

Also im trying to use a lectin on an unfixed sample.

Any and all suggestions welcome

350um THICK sections, Lectin histochemistry NOT immunohisto!
« on: October 07, 2009, 12:09:02 PM »

Offline excalibur

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 1109
    • http://www.excaliburpathology.com
Re: 350um THICK sections, Lectin histochemistry NOT immunohisto!
« Reply #1 on: October 07, 2009, 05:50:55 PM »
It should have come with a datasheet as to a recommended dilution. Vector is usually very good about sending datasheets. If you did not get one, it is on their website with the product info.

350 micron sections are unusually thick. You will need to incubate longer than an hour or do free floating staining.
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Offline CanuckPhD

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 296
Re: 350um THICK sections, Lectin histochemistry NOT immunohisto!
« Reply #2 on: October 08, 2009, 02:33:53 AM »
Is there any reason that you need to use such thick unfixed sections? I think you can get around the issues of the thickness by using Triton-X and long incubations (48h at least). However, if you do all of those things your unfixed section will be completely destroyed (IMHO).

If you are using WFA to look for perineural nets there are reports in the literature of using PFA fixed thick sections from both humans and rodents, so it should be possible. I have used PFA postfixed sections (10um) to look at perineural nets in rats and had good luck.

 

Re: 350um THICK sections, Lectin histochemistry NOT immunohisto!
« Reply #2 on: October 08, 2009, 02:33:53 AM »